Day 3 :
- Gene Therapy, Cell and Gene Therapy Products, Diabetes for Gene Therapy, Viral Gene Therapy, Genetically Inherited diseases
Location: Lone Star West
Kiev regional p/n hospital, Germany
University of California, USA
Kunwoo Lee is getting his PhD in May 2016. He has published more than 8 papers in reputed journals
The CRISPR/Cas9 technology has the potential to achieve therapeutic genome editing with high specificity and efficiency. However, the applications of Cas9 are limited by the lack of delivery vehicles that can simultaneously transduce proteins, guide RNA, and donor oligonucleotides into cells for genome editing. In this report we demonstrate that gold nanoparticles conjugated with DNA and assembled with endosomal disruptive polymers, can deliver Cas9 protein, guide RNA (gRNA) and donor DNA into various cells, including stem cells, and induce homology directed repair (HDR) with exceptional efficiency. Cellular treatment of the vehicle resulted in efficient cellular uptake. HDR experiments with donor ssDNA with restriction enzyme site showed that 15 nm gold nanoparticles complexed with Cas9 RNP can induce HDR in human embryonic stem (hES) and human induced pluripotent stem (hiPS) cells, with a significantly high efficiency of 5-6%. Compared to the conventional nucleofection method that resulted in low HDR efficiency in stem cells with high toxicity, GNP platform can efficiently deliver Cas9 RNP without cell detachment from culture plate and with significantly low toxicity. In addition, we demonstrate that gold nanoparticles can deliver Cas9 RNP in vivo to muscle tissue in mice. Intramuscular injection in mice resulted in significant uptake of fluorescently labeled Cas9 RNP in muscle tissue. We have developed a new delivery vehicle that can deliver both Cas9 RNP and donor DNA together for efficient HDR. Efficient editing of endogenous genes was achieved in many mouse and human cells
Kiev regional p/n hospital, Ukraine
Ponizovskiy M R has graduated from N.I.Pirogov Vinitsa National Medical University. He worked manager of the hospital in Sum district of Ukraine and then as a Doctor of therapeutic department in the Kiev regional hospital. Then he has graduated from Kiev national Institute of post diploma education and Lvov National Medical University, faculties of laboratory clinical diagnostics, laboratory clinical biochemistry, laboratory toxicology. In 1972 – 1990, he worked as the head of clinical and biochemical laboratory of the First Kiev regional hospital, and I have obtained Degree PhD from 1990. In 1990 – 2002, he worked as the head of toxicological and biochemical Laboratory of the Kiev regional p/n hospital. He has the scientific degree PhD and the clinical degree of the highest category doctor. In Germany, he is the member of the Society of Inventors: he has the German protection of the invention [Gebrauchsmusters] of the new method of blood cells separation and allocation, and this method is examined in German Patent Department for reception of the German Patent.
The significant separate biochemical discoveries of pro-/anti-apoptotic processes, pro-/anti-autophagy processes, pro-/antiproliferative processes in norm and pathology gave possibility to propose the common concept all of these processes mechanism: Interactions between all cells of an organism occur due to remote reactions across distance as the results of cellular capacitors operations via production of resonance waves that maintain common stability of Internal Energy and Internal Medium both in cells and in an organism. Intracellular balances catabolic and anabolic processes interconnect with extracellular balances catabolic and anabolic processes promoting as maintenance stability Internal Medium and Internal Energy of cells as well as cell development in norm. On the contrary the violations interconnections of intracellular balances catabolic & anabolic processes with extracellular balances catabolic & anabolic processes promote pathologic processes. The excessive shifts balance catabolic & anabolic processes into catabolic processes leads to inflammatory or infectious processes expressions. The excessive shifts balance catabolic & anabolic processes into anabolic processes leads to cancer diseases expressions which are characterized by irrepressible proliferative processes due to abundance of ROS/H2O2/free radicals operation resulting in excessive 2nDNA reaction. Studying cellular cycle mechanisms in norm and pathology it was explained the mechanisms of maintenance stability cellular Internal Medium and Internal Energy due to interactions between nuclear function and mitochondrial function, which, on the one hand, link with the stability Internal Energy and Internal Medium of an organism, and, on the other hand, the violation normal interactions between nuclear function and mitochondrial function leads to quasi-stationary pathologic states of cellular Internal Energy and Internal Medium. Besides it was elucidated the violations mechanisms of maintenance stability cellular Internal Medium and cellular Internal Energy in cancer tissue in comparison with the violations mechanisms of maintenance stability cellular Internal Medium and cellular Internal Energy in inflammatory processes. All of it gave possibility to eliminate a lot of doubts and/or queries which were expressed by the authors of the some experiments
Mohamed M Kamal finished his PhD from Faculty of Pharmacy, Ain Shams University, Cairo, Egypt in 2011 on glioblastoma cancer stem cell biology in Genetics Department, M.D. Anderson Cancer Center, Houston, Texas from 2008 through 2010 as part of scholarship from Egyptian Government. In 2014, he got a Fulbright Postdoctoral fellowship in M.D. Anderson Cancer Center, Houston, Texas for nine months. He published 2 papers in Stem Cells and Journal of Neuroscience as a first coauthor. Also, he has a paper published in Regenerative Medicine and a couple of papers under revision in stem cell biology and cancer fields
Background: The number of patients suffering from diabetes mellitus (DM) is growing in an alarming rate. Now, cell therapy treatment options for DM are under extensive study. Interestingly, umbilical cord (UC) has been proved to be a good source of stem cells, either from cord blood (UCB-MSCs) or Wharton’s jelly (WJ-MSCs).Nowadays, with the evolving interest in UC stem cells banking, we thought to investigate the difference between these 2 important banking sources and to compare their differentiation potentials towards insulin producing cells (IPCs) in vitro and in vivo. Methods: Both UCB-MSCs and WJ-MSCs were isolated and expanded for several passages. Afterwards, both cell types were induced to differentiate into IPCs, then the differentiated cells were assessed by determining the expression of stem cell markers, together with key β-cells markers using qRT-PCR, and functionally by measuring glucose stimulated insulin secretion. Both UCB-MSCs and WJMSCs were transplanted in the tail veins of streptozotocin (STZ) induced diabetic rats. Blood glucose levels and body weights were monitored 2 months post transplantation. Results & Conclusion: WJ appeared to be a much more homogenous and potential source for MSCs as compared to UCB. Interestingly, both UCB-MSCs and WJ-MSCs were successfully differentiated to IPCs. Yet, the resulting IPCs from WJ-MSCs were to a limited extent functioning better than those obtained from UCB-MSCs in vitro. Moreover, WJ-MSCs managed to better control hyperglycemia in STZ induced diabetic rats. Our results indicate that WJ could represent a potential source of cells in the field of DM cell therapy.
King Saud University, Saudi Arabia
Abdullah completed his PH.D and presently working as an assistant professor in King Saud University, Saudi Arabia
Aim: The aim of this study was to investigate the incidence of hepatitis A, B and C in Kingdom of Saudi Arabia, and to determine which age group is most commonly infected by which one of the hepatitis viruses A, B or C. Background: The epidemiology of viral hepatitis in Saudi Arabia has undergone major changes, concurrent with major socioeconomicdevelopments over the last two to three decades.This disease represents a major public health problem in Saudi Arabia resulting in the need for considerable healthcare resources. Method: A retrospective analysis of the reported cases of viral hepatitis was conducted based on the reports of The Ministry of Health in Saudi Arabia about Hepatitis A, B and C cases in all the regions of Saudi Arabia from 2006 to 2010. Results: Our study demonstrated that the incidence of viral hepatitis showed a decreasing pattern over the study period, except for the incidence of hepatitis B in Saudis that showed a small increase. Of hepatitis A, B, and C, HBV was the most predominant type of hepatitis, accounting for (53%) of the cases, followed by HCV (30%) and HAV (17%). HAV infection predominates in children (5–14 years) with 60% of HAV cases, HBV in young adults (15–44 years) with 69% of HBV cases, and HCV in older adults (>45 years) with 59% of cases (p < 0.01). Conclusion: Despite significant changes in the incidence of viral hepatitis A, B and C, it remains a major public health problem in Saudi Arabia; however, it showed a significant decline in the last two decades that could be attributed to the nationwide vaccination programs and the improved health facilities, but further investigations and better health controls are needed to control the increase in HBV incidence
B.P. Koirala Institute of Health Sciences (BPKIHS), Nepal
Deepak Kumar Roy is a postgraduate resident in department of conservative dentistry and Endodontics at BPKIHS, Dharan, Nepal. He has completed his graduation in dentistry from BPKIHS, Dharan Nepal. He has deep interest in research works and has multiple national and international publications. He has worked in genetics, cleft lip and palate, dental caries, psychological aspects and involved in innovative works. Recently, he has successfully completed research on influence of genetics on dental caries.
Dental caries is multi factorial in origin. Various etiologic factors are responsible for the causation of disease process. There is an emerging evidence for a genetic component in caries susceptibility. Studies in human have suggested that variation in enamel formation genes may contribute to caries development. Functional polymorphisms in the Matrixmetalloproteinase (MMP) genes have been attributed to enamel development and caries pathogenesis. The objective of this study was to investigate the relationship of MMP-13 (rs2252070) to dental caries. The sample consisted of 60 subjects. Oral hygiene status was accessed by simplified Oral hygiene Index (OHI-S). Dental caries was assessed according to the criteria recommended by the World Health Organization guidelines using the DMFT index. The subjects were divided into 4 groups (15 in each group). Group A: subjects having poor oral hygiene and absence of dental caries. Group B: Poor oral hygiene and presence of dental caries. Group C: good oral hygiene and absence of dental caries. Group D: good oral hygiene and presence of dental caries. Blood samples (1.5 ml) were obtained from the patients followed by isolation of Genomic DNA, Polymerase chain reaction test and digestion with enzyme Bsr-I. Result indicated that due to the single nucleotide polymorphism of MMP-13(rs2252070) the patients were suffering from dental caries despite having good oral hygiene. The role of MMP-13 was not evident for age and gender with dental caries
Mansoura University School of Medicine, Egypt
Heba Mosaad completed her PH.D and presently working in Mansoura University School of Medicine, Egypt
Background: Juvenile idiopathic arthritis (JIA) is the most common rheumatic disease of the childhood with a high risk of disability in Egyptian children. JIA has many genetic factors affecting its pathogenesis including CD226 and CD40 genes. These genetic factors vary in different races proved by previous studies on European and Chinese populations. The association between JIA and CD226 and CD40 is yet to be evaluated in non-European populations including Egypt. So we studied the association of CD226 rs1883832 (-1C>T) and CD40 rs1883832 (-1C>T) gene polymorphism and disease susceptibility and severity of in an Egyptian cohort. Subject & Methods: In this case control study we recruited 79 Egyptian children with JIA and 93 healthy controls. We studied CD226 rs763361 (C>T) using the tetra amplification refractory mutation system - polymerase chain reaction assay (ARMS-PCR) for detection of polymorphism while for CD40 rs1883832 (-1C>T) we used restriction fragment length polymorphism (RFLP). Results: The statistical results showed that the rs763361 (C>T) SNP in the CD226 gene is significantly associated with JIA group as regard to TT genotypes (p= 0.0001). The frequency of the T allele was significantly higher in JIA patients in comparison with the control group (p= 0.0001). Also this allele was significantly higher in patients with moderate and sever JIA when compared to controls (p=0.003). This allele correlated to the disease severity (OR=2.4). Study of CD40 rs1883832 (-1C>T) showed that the distribution of the C allele was significantly higher in JIA patients (p=0.003). Also it was significantly higher in patients with moderate and sever JIA when compared to controls (p=0.01). Conclusion: These results demonstrate a genetic association between the CD226 and CD40 gene polymorphism and JIA with an impact on disease severity in an Egyptian cohort